Evaluation and applications of optical cell density probes in mammalian cell bioreactors

On-line optical cell density probes were implemented to continuously monitor the cell densities in mammalian cell bioreactor and to achieve advanced bioreactor controls. We tested cell density probes from six manufacturers in high cell density bioreactors. When externally calibrated, Aquasant and Ingold backscattering probes produced the most linear probe responses (PR) versus cell density (CD), followed by the ASR and Cerex laser probes. Monitek and Wedgewood transmission probes had lower resolutions. All probes were tested in two murine hybridoma fermentations. Cell densities varied between 1 x 10(6) cells/mL to 20 x 10(6) cells/mL and the bioreactors were operated for 5 to 7 weeks. For our bioreactors, Aquasant, Ingold, ASR, Wedgewood, and Monitek probes gave satisfactory responses. Little fouling was observed with any probe at the end of 2 weeks. Fouling was a possibility after 3 weeks in one bioreactor but its effect can be easily corrected. Cell density control and specific perfusion control of bioreactors based on the Aquasant probe were achieved. Implementation of cell density probe based perfusion control, instead of "step perfusion adjustments" based on manual hemacytometer control, will result in smoother operation, healthier cultures, increased medium delivery efficiency, and reduced operational excursions. (c) 1995 John Wiley & Sons, Inc.     

Impacts of the pesticides imidacloprid,propargite, and pymetrozine on <Emphasis Type="Italic">Chrysoperla carnea</Emphasis> (Stephens) (Neuroptera: Chrysopidae): IOBC and life table assays

The effects of imidacloprid, propargite, and pymetrozine on the common green lacewing, Chrysoperla carnea (Stephens) were investigated in the laboratory, using the IOBC-system and the life table response experiment. Residual glass plate bioassays were carried out using two-day-old larvae at the Iranian maximum field recommended rate of each commonly used pesticide. All three tested pesticides produced significant adverse effects on pre-imaginal survival (p < 0.01). Imidacloprid had no significant effect on fecundity, but propargite and pymetrozin caused significant reductions (p < 0.05). According to the IOBC classification, imidacloprid was found to be harmless (E = 27.44%); propargite (E = 49.78%) and pymetrozine (E = 66.9%) were slightly harmful. Life table assays revealed that imidacloprid and propargite had no significant effects on the intrinsic rate of natural increase, while pymetrozine caused a 34% reduction in r _m value (p < 0.05). Propargite was non-toxic to C. carnea under the tested conditions. In the case of imidacloprid, using a glass plate as test substrate led to␣underestimation of its effects. The life table assay showed more adverse effects of pymetrozine than the IOBC method.     

PAX6 Regulates Melanogenesis in the Retinal Pigmented Epithelium through Feed-Forward Regulatory Interactions with MITF

During organogenesis, PAX6 is required for establishment of various progenitor subtypes within the central nervous system, eye and pancreas. PAX6 expression is maintained in a variety of cell types within each organ, although its role in each lineage and how it acquires cell-specific activity remain elusive. Herein, we aimed to determine the roles and the hierarchical organization of the PAX6-dependent gene regulatory network during the differentiation of the retinal pigmented epithelium (RPE). Somatic mutagenesis of Pax6 in the differentiating RPE revealed that PAX6 functions in a feed-forward regulatory loop with MITF during onset of melanogenesis. PAX6 both controls the expression of an RPE isoform of Mitf and synergizes with MITF to activate expression of genes involved in pigment biogenesis. This study exemplifies how one kernel gene pivotal in organ formation accomplishes a lineage-specific role during terminal differentiation of a single lineage.     

Effect of Histamine H2 Receptor Antagonists on the Secretion of Cerebrospinal Fluid in the Cat

Following a recent report that epithelial cells of the choroid plexus possess histamine H2 receptors, the effect of cimetidine and ranitidine, histamine H2 receptor antagonists, on the secretion and electrolyte content of CSF was examined. Fifty cats were divided into one control (n = 6) and six experimental groups. CSF was collected by puncture of the cisterna magna following pentobarbital anesthesia, and its volume, concentrations of Na+, K+, Cl-, and pH were determined. Cimetidine or ranitidine (50, 20, or 10 mg/kg) was injected intravenously 2 h after the start of the test, and their concentrations were measured in hourly blood samples and in 30-min aliquots of CSF in the 50 mg/kg experimental groups. Whereas the secretion of CSF did not change over 6 h in the control group, it decreased significantly by 30-60 min after injection of cimetidine or ranitidine and remained low for the following 6 1/2 h in all experimental groups except the 10-mg ranitidine group. Peak cimetidine and ranitidine concentrations in CSF in the 50-mg experimental groups were noted 60 and 90 min, respectively, after intravenous injection. CSF electrolyte concentrations and pH did not change during the test in any group. We conclude that intravenous cimetidine or ranitidine can significantly reduce CSF secretion in the cat, possibly by competitive inhibition of the histamine effect on H2 receptors located on the choroid plexus epithelial cell, or by a direct effect on the capillaries of the choroid plexus.     

NATURAL SELECTION OF ALLOZYME POLYMORPHISMS: A MICROSITE TEST REVEALING ECOLOGICAL GENETIC DIFFERENTIATION IN WILD BARLEY

Allozymic variation in proteins encoded by 25 loci was analyzed electrophoretically in 1982 and 1983 in 356 individual plants from a dense population of wild barley, Hordeum spontaneum, the progenitor of cultivated barley. The test involved six microniches organized in a mosaic pattern in the open Tabor oak forest at Neve Ya'ar, Israel. The microniches were i) sun-soil, ii) sun-rock, iii) shade-soil, iv) shade-rock, and the contact zones: v) soil periphery of the sun-rock microniche, and vi) soil periphery of the shade-rock microniche. Discriminant analysis indicated significant multilocus allozymic differentiation between the microniches. Our results suggest that allozyme polymorphisms in wild barley are at least partly adaptive and differentiate predominately by microniche ecological selection, rather than by stochastic processes and/or neutrality of allozymic variants.     

Renal structural flexibility in response to environmental water stress in feral hogs

Several morphological characteristics of the kidney were studied to determine the degree of acclimatization that may occur in three groups of feral hogs raised under different environmental conditions. Two groups of hogs were living in the wild, while another was raised in captivity for three generations and was directly descended from one of the wild-living groups. The two groups of wild hogs were living under two different types of water stress conditions. One group experienced periodic drought, and the other ate a high salt diet. The captive hogs were given food and water ad libitum. The captive-raised hogs had significantly lower relative medullary thickness (RMT) and relative medullary area (RMA) values (RMT of 2.35; RMA of 0.35) than either group of hogs living in the wild (RMT of 2.70 and 2.69; RMA of 0.41 and 0.44). Since the feral hogs living in the wild were exposed to a higher degree of water stress than the captive-raised hogs, it was concluded that the differences in observed kidney structure were due to acclimatization.     

Development and fecundity of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) on different soybean cultivars

The life history of Spodoptera exigua was studied under laboratory conditions at temperature of 25°C ± 1°C, relative humidity (RH) of 60% ± 5% and a photoperiod of 16:8 h (Light:Dark) on nine soybean (Glycine max L.) cultivars: 032, 033, Hill, M4, M7, M9, M11, TMS and Zan. Larval and pupal development times were longest on M7 cultivar (14.09 ± 0.078 and 7.78 ± 0.090 days) and shortest on 033 cultivar (11.98 ± 0.054 and 7.033 ± 0.090 days). The longest and shortest female and male longevities were observed on Hill and Zan cultivars, respectively. The results showed that the whole life span varied from 32.80 ± 0.89 to 37 ± 0.98 days for females and from 33.3 ± 0.66 to 38.74 ± 0.95 days for males. The highest daily fecundity was obtained on M9 (102. 38 eggs), and the lowest on Hill (75.87 eggs). Mean total fecundity per female during adult life was highest on 033 (1156.20 eggs) and lowest on M7 cultivars (841.68 eggs). Clustering analysis of the biological parameters of S. exigua confirmed that 033 and Hill cultivars were the susceptible and the resistant cultivars, respectively.     

Melanoma‐associated leukoderma – immunology in black and white?

Melanoma is an ‘immunogenic tumor’, often highly infiltrated with lymphocytes, which are capable of inducing regression of the primary tumor. The commonly observed phenomenon of regression suggests substantial cross‐talk between immune cells and transformed melanocytes. An immune response to melanocyte differentiation antigens common to transformed and normal melanocytes manifests clinically at distant sites as melanoma‐associated vitiligo or halo nevi. Despite similar antigenic targets, the pathogenesis and prognosis differ between the different melanoma‐associated leukodermas. Understanding immunologic cross‐talk between melanocytes and the immune system will aid the development of approaches to combat melanoma.